Laboratory of Molecular NeuroTherapeutics

PI: Tsuneya Ikezu, M.D., Ph.D.

Figure 1. Imaging of Aβ aggregation in vitro.  4D imaging of Aβ aggregation.  0.1% QDAβ(6)-containing Aβ42 (final concentration 100 µM) were incubated in a 96-well glass bottom plate at 37 ºC with controlled humidity and CO2 concentration, and observed by confocal microscopy every 30 min over 20 h using a 488 nm excitation laser (20% power) and a 60x oil objective lens (PLoS One 4:e8492).
Figure 1. Imaging of Aβ aggregation in vitro. 4D imaging of Aβ aggregation. 0.1% QDAβ(6)-containing Aβ42 (final concentration 100 µM) were incubated in a 96-well glass bottom plate at 37 ºC with controlled humidity and CO2 concentration, and observed by confocal microscopy every 30 min over 20 h using a 488 nm excitation laser (20% power) and a 60x oil objective lens (PLoS One 4:e8492).

Research Interests:

The Laboratory of Molecular NeuroTherapeutics mainly focuses on neuroimmune cell-mediated regulations of neuronal function, neurogenesis, neuroinflammation, and neurodegeneration. In particular, we are interested in how the innate immune-related molecules in the central nervous system (CNS) influences the pathology and progression of select neurodegenerative disorders e.g. Alzheimer’s Disease, Frontotemporal Dementia, NeuroAIDS, and recently Chronic Traumatic Encephalopathy.

Figure 2. 3D reconstruction of amyloid plaque deposition and atro/microgliosis in APP mouse brain. Cortical section of Tg2576 (14 months of age) was stained by thioflavin-S (compact amyloid plaque, purple), anti-IBA1 rabbit polyclonal (microglia, red), and anti-GFAP mouse monoclonal (astrocytes, green), and imaged using a laser scanning confocal microscope (Nikon SweptField). Z-stack images were 3D-deconvolved and 3D-reconstructed using image processing software (AutoQuant). Original magnification, X400.
Figure 2. 3D reconstruction of amyloid plaque deposition and astro/microgliosis in APP mouse brain. Cortical section of Tg2576 (14 months of age) was stained by thioflavin-S (compact amyloid plaque, purple), anti-IBA1 rabbit polyclonal (microglia, red), and anti-GFAP mouse monoclonal (astrocytes, green), and imaged using a laser scanning confocal microscope (Nikon SweptField). Z-stack images were 3D-deconvolved and 3D-reconstructed using image processing software (AutoQuant). Original magnification, X400.

Current studies include pharmacological means to suppress the innate immune response in the CNS with the goal of enhancing neuronal protection, and minimizing collateral damage from an activated immune response in the CNS.  Specifically, we are investigating the role of the signaling cytokine IL-4/10/CD200 and its potential anti-inflammatory and neuroprotective role between microglia and both neurons and astrocytes during active CNS inflammation.  Further, we are exploring IL-4/10/CD200 as a potential therapeutic in chronic inflammatory states within the CNS, such as seen in Alzheimer’s disease, to minimize neuronal cell loss.

A second focus of our lab is the investigation of tau-tubulin kinases (TTBK1 and 2) and their role in protein phosporylation leading to the formation of tau and alpha-synuclein aggregates within neurons, a key pathology observed in AD, Amyotrophic Lateral Sclerosis, Spinocerebellar Ataxia, and Frontotemporal Dementia .  We are currently performing a cutting-edge drug discovery program to identify potential inhibitors to TTBK1 with the goal of identifying potential new therapeutic(s) that could prevent tangle formation within neurons in these neurodegenerative states.  We primarily use novel gene-targeted/transgenic mouse models to characterize our studies by employing a mix of assays including: standard biochemical assays, gene expression analyses, tissue cultures from both primary cells and cell lines, immunohistochemistry, gene-delivery vectors, and animal behavioral testing.

Figure 3. AAV1/2 hybrid gene delivery of IL-4 and IL-10 in hippocampus enhances neurogenesis in SGZ of APP+PS1 mice. Immunofluorescence of BrdU (green) and NeuN (red) in the dentate gyrus of non-Tg or APP+PS1 mice injected with AAV-GFP, AAV-IL-4, or AAV-IL-10 at 3 months of age and ip injected with BrdU 3 weeks prior to the euthanasia at 8 months of age. Arrows indicate NeuN+BrdU+ new neurons in SGZ.
Figure 3. AAV1/2 hybrid gene delivery of IL-4 and IL-10 in hippocampus enhances neurogenesis in SGZ of APP+PS1 mice. Immunofluorescence of BrdU (green) and NeuN (red) in the dentate gyrus of non-Tg or APP+PS1 mice injected with AAV-GFP, AAV-IL-4, or AAV-IL-10 at 3 months of age and ip injected with BrdU 3 weeks prior to the euthanasia at 8 months of age. Arrows indicate NeuN+BrdU+ new neurons in SGZ.